基于多肽水凝胶材料的肿瘤类器官培养体系的建立

Development of a tumor organoid culture system with peptide-based hydrogels

摘要:

多肽水凝胶是一类具有特殊网络结构的高分子材料，因其性质稳定、生物相容性良好等特点被广泛应用于生物医药领域。环境响应型自组装多肽水凝胶在环境改变时发生响应，多肽自组装形成纳米纤维网状结构，更好地模拟了细胞外基质和细胞生长微环境，可用于细胞的3D培养和类器官培养。为了建立CulX Ⅱ多肽水凝胶材料的肿瘤类器官培养体系，本研究选用了Panc-1、U87、H358细胞，采用半球体的培养方式，通过CulXⅡ多肽水凝胶材料包裹肿瘤细胞在24孔板中培养15 d。胰腺癌肿瘤类器官呈现出3D立体球体的形态，类器官大小随培养时间的延长而增大，最终直径大小为150−300 μm。肿瘤类器官数量较多、大小不一、细胞活力较好、边缘轮廓清晰、形态较好，培养比较成功。本研究在CulX Ⅱ多肽水凝胶材料基础上建立了肿瘤类器官模型，为研究肿瘤发病机制、新药研发、肿瘤抑制提供了研究模型。

关键词:自组装;多肽水凝胶;类器官;3D培养;肿瘤

Abstract:

Peptide-based hydrogel, the polymer materials with a special network structure, are widely used in various fields of biomedicine due to their stable properties and biocompatibility. Environment-responsive self-assembled peptide aqueous solutions can respond to environment changes by the self-assembly of peptides into nanofiber networks. Peptide-based hydrogels well simulate the extracellular matrix and cell growth microenvironment, being suitable for 3D cell culture and organoid culture. To establish a tumor organoid culture system with peptide-based hydrogels, we cultured Panc-1, U87, and H358 cells in a 3D spherical manner using CulX Ⅱ peptide-based hydrogels in 24-well plates for 15 days. The organoids showed a 3D spherical shape, and their sizes increased with the extension of the culture time, with a final diameter ranging from 150 to 300 μm. The organoids had a large number, varying sizes, good cell viability, clear edges, and a good shape, which indicated successful organoid construction. The tumor organoid culture system established in this study with CulX Ⅱ peptide-based hydrogels provides a model for studying tumor pathogenesis, drug development, and tumor suppression.

Key words:self-assembly;peptide-based hydrogel;organoid;3D culture;tumor